Type of Submission
Poster
Keywords
Tetrahymena, CRISPR, oncohistone, H3K27M
Proposal
CRISPR-Cas9 editing is a powerful tool for making genome changes in organisms. The free-living ciliated protozoan, Tetrahymena thermophila, has been used for decades as a model system for eukaryotic genetic and epigenetic phenomena. Because of this, there is a great deal of interest in adapting the CRISPR/Cas9 system to Tetrahymena. Plasmids containing the gene for Cas9 nuclease are currently available for transfection into Tetrahymena. However, the transfection protocols which are currently used in this organism, electroporation or biolistic transformation using a gene gun, are both expensive and inefficient. We are attempting to use liposomes in order to transfect Tetrahymena with the Cas9 nuclease. Our hope is to develop a protocol that would allow for a more efficient, less expensive transfection platform, facilitating further development of CRISPR-Cas9 editing in this system.
Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.
Can Liposomes be Used to Transfect Tetrahymena Thermophila for CRISPR/Cas9 Studies?
CRISPR-Cas9 editing is a powerful tool for making genome changes in organisms. The free-living ciliated protozoan, Tetrahymena thermophila, has been used for decades as a model system for eukaryotic genetic and epigenetic phenomena. Because of this, there is a great deal of interest in adapting the CRISPR/Cas9 system to Tetrahymena. Plasmids containing the gene for Cas9 nuclease are currently available for transfection into Tetrahymena. However, the transfection protocols which are currently used in this organism, electroporation or biolistic transformation using a gene gun, are both expensive and inefficient. We are attempting to use liposomes in order to transfect Tetrahymena with the Cas9 nuclease. Our hope is to develop a protocol that would allow for a more efficient, less expensive transfection platform, facilitating further development of CRISPR-Cas9 editing in this system.
